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Polymorphism analysis by nucleic acid structure probing with structure-bridging oligonucleotides

  • US 6,210,880 B1
  • Filed: 09/19/1997
  • Issued: 04/03/2001
  • Est. Priority Date: 09/19/1997
  • Status: Expired due to Term
First Claim
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1. A method for capturing bridging oligonucleotides, comprising:

  • a) providing;

    i) a first folded target having a nucleic acid sequence comprising first and second portions, said first and second portions each comprising one or more double stranded regions and one or more single stranded regions;

    ii) a second folded target having a nucleic acid sequence comprising a first portion that is identical to said first portion of said first folded target and a second portion that differs from said second portion of said first folded target because of a variation in nucleic acid sequence relative to said first folded target, said first and second portions each comprising one or more double stranded regions and one or more single stranded regions;

    iii) first and second bridging oligonucleotides said first bridging oligonucleotide complementary to said first portion of said first and second folded targets and said second bridging oligonucleotide complementary to said second portion of said first and second folded targets; and

    iv) a solid support comprising first, second, third and fourth capture zones, each zone capable of capturing and immobilizing said first and second bridging oligonucleotides;

    b) contacting said first folded target with said first bridging oligonucleotide under conditions such that said first bridging oligonucleotide binds to said first folded target to form a probe/folded target complex in a first mixture;

    c) contacting said first folded target with said second bridging oligonucleotide under conditions such that said second bridging oligonucleotide binds to said first folded target to form a probe/folded target complex in a second mixture;

    d) contacting said second folded target with said first bridging oligonucleotide to form a third mixture;

    e) contacting said second folded target with said second bridging oligonucleotide to form fourth mixture; and

    f) adding said first, second, third and fourth mixtures to said first, second, third and fourth capture zones of said solid support, respectively, under conditions such that said first and second bridging oligonucleotides are captured and immobilized.

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